scholarly journals Current Status of Matrix-Assisted Laser Desorption/Ionization–Time-of-Flight Mass Spectrometry (MALDI-TOF MS) in Clinical Diagnostic Microbiology

Molecules ◽  
2020 ◽  
Vol 25 (20) ◽  
pp. 4775
Author(s):  
Sachio Tsuchida ◽  
Hiroshi Umemura ◽  
Tomohiro Nakayama

Mass spectrometry (MS), a core technology for proteomics and metabolomics, is currently being developed for clinical applications. The identification of microorganisms in clinical samples using matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry (MALDI-TOF MS) is a representative MS-based proteomics application that is relevant to daily clinical practice. This technology has the advantages of convenience, speed, and accuracy when compared with conventional biochemical methods. MALDI-TOF MS can shorten the time used for microbial identification by about 1 day in routine workflows. Sample preparation from microbial colonies has been improved, increasing the accuracy and speed of identification. MALDI-TOF MS is also used for testing blood, cerebrospinal fluid, and urine, because it can directly identify the microorganisms in these liquid samples without prior culture or subculture. Thus, MALDI-TOF MS has the potential to improve patient prognosis and decrease the length of hospitalization and is therefore currently considered an essential tool in clinical microbiology. Furthermore, MALDI-TOF MS is currently being combined with other technologies, such as flow cytometry, to expand the scope of clinical applications.

Author(s):  
Rosa M Gomila ◽  
Gabriel Martorell ◽  
Pablo A Fraile-Ribot ◽  
Antonio Doménech-Sánchez ◽  
Miguel Albertí ◽  
...  

Abstract Background Classification and early detection of severe COVID-19 patients is required to establish an effective treatment. We tested the utility of matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to classify and predict the severity of COVID-19. Methods We used MALDI-TOF MS to analyse the serum peptidome from 72 COVID-19 patients (training cohort), clinically classified as mild (28), severe (23) and critical (21), and 20 healthy controls. The resulting matrix of peak intensities was used for Machine Learning (ML) approaches to classify and predict COVID-19 severity of 22 independent patients (validation cohort). Finally, we analysed all sera by liquid chromatography mass spectrometry (LC MS/MS) to identify the most relevant proteins associated to disease severity. Results We found a clear variability of the serum peptidome profile depending on COVID-19 severity. Forty-two peaks exhibited a log fold change ≥ 1 and 17 were significantly different and at least four-fold more intense in the set of critical patients than in the mild ones. ML approach classified clinical stable patients according to their severity with a 100% of accuracy and predicted correctly the evolution of the non-stable patients in all cases. LC MS/MS identified five proteins that were significantly upregulated in the critical patients. They included the serum amyloid protein A2, which probably yielded the most intense peak detected by MALDI-TOF MS. Conclusion We demonstrated the potential of the MALDI-TOF MS as a bench to bedside technology to aid clinicians in their decisions on COVID-19 patients.


2012 ◽  
Vol 90 (9) ◽  
pp. 1117-1127 ◽  
Author(s):  
E.L. Pannkuk ◽  
D.F. Gilmore ◽  
B.J. Savary ◽  
T.S. Risch

Lipids secreted from sebaceous glands in bat integument may play a role in determining host pathogenicity by the fungus Geomyces destructans in white-nose syndrome (WNS). To investigate this, we have determined the triacylglycerol (TAG) profiles for three bat species: eastern red bats ( Lasiurus borealis (Müller, 1776)), evening bats ( Nycticeius humeralis (Rafinesque, 1818)), and big brown bats ( Eptesicus fuscus (Beauvois, 1796)). Neutral lipids extracted from the hair and wing tissue were fractionated by preparative thin-layer chromatography (TLC) into four major lipid bands corresponding to cholesterol, free fatty acids (FFAs), TAGs, and sterol–wax esters. Densitometry showed higher proportions of TAGs in hair than from wing tissue. TAG bands were recovered and analyzed by matrix-assisted laser desorption–ionization time-of-flight mass spectrometry (MALDI–TOF MS). Mass spectra showed sodiated TAG species with variable fatty acyl (FA) moieties range from m/z 715.6–911.8. High intensity ion peaks were consistent with 16:0 and 18:1 as dominant FA moieties, and these were identified as palmitic and oleic acids, respectively, by liquid chromatography electrospray ionization mass spectrometry. We determined significant differences in TAG profiles between three bat species by MALDI–TOF MS, providing the first description of integumentary lipids in bats. In this study, we performed the first TAG profiling of bats, which suggest such profiles may be species-specific in bats.


HU Revista ◽  
2018 ◽  
Vol 43 (3) ◽  
pp. 285-294
Author(s):  
Flávia Lúcia Piffano Costa Pellegrino ◽  
Thiago Pavoni Gomes Chagas ◽  
Maria Silvana Alves ◽  
Ana Paula D’Alincourt Carvalho-Assef ◽  
Alexander Chapeaurouge ◽  
...  

Among its innumerous applications in Bacteriology, the Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) technique is evolving as a powerful tool for bacterial identification and antimicrobial resistance investigation. Publications have evaluated the MALDI-TOF MS performance in the identification of a series of bacterial pathogens, including the most common severe infectious agents, emergent pathogens involved with outbreaks of healthcare-associated infections, rare pathogens, and those whose isolation in culture media is difficult. As compared to conventional methods of bacterial identification, MALDI-TOF MS has proven to be a fast, accurate and cost-effective technique. Currently, MALDI-TOF MS has been used in antimicrobial resistance studies, since it has shown to be an efficient tool in detecting specific resistance mechanisms in bacteria, such as beta-lactamases production, for example. Here, we describe the advances in this growing field of mass spectrometry applied to Bacteriology, including Brazilian contributions.


2017 ◽  
Vol 55 (5) ◽  
pp. 1437-1445 ◽  
Author(s):  
Maya Beganovic ◽  
Michael Costello ◽  
Sarah M. Wieczorkiewicz

ABSTRACT Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) decreases the time to organism identification and improves clinical and financial outcomes. The purpose of this study was to evaluate the impact of MALDI-TOF MS alone versus MALDI-TOF MS combined with real-time, pharmacist-driven, antimicrobial stewardship (AMS) intervention on patient outcomes. This single-center, pre-post, quasiexperimental study evaluated hospitalized patients with positive blood cultures identified via MALDI-TOF MS combined with prospective AMS intervention compared to a control cohort with MALDI-TOF MS identification without AMS intervention. AMS intervention included: real-time MALDI-TOF MS pharmacist notification and prospective AMS provider feedback. The primary outcome was the time to optimal therapy (TTOT). A total of 252 blood cultures, 126 in each group, were included in the final analysis. MALDI-TOF MS plus AMS intervention significantly reduced the overall TTOT (75.17 versus 43.06 h; P < 0.001), the Gram-positive contaminant TTOT (48.21 versus 11.75 h; P < 0.001), the Gram-negative infection (GNI) TTOT (71.83 versus 35.98 h; P < 0.001), and the overall hospital length of stay (LOS; 15.03 versus 9.02 days; P = 0.021). The TTOT for Gram-positive infection (GPI) was improved (64.04 versus 41.61 h; P = 0.082). For GPI, the hospital LOS (14.64 versus 10.31 days; P = 0.002) and length of antimicrobial therapy 24.30 versus 18.97 days; P = 0.018) were reduced. For GNI, the time to microbiologic clearance (51.13 versus 34.51 h; P < 0.001), the hospital LOS (15.40 versus 7.90 days; P = 0.027), and the intensive care unit LOS (5.55 versus 1.19 days; P = 0.035) were reduced. To achieve optimal outcomes, rapid identification with MALDI-TOF MS combined with real-time AMS intervention is more impactful than MALDI-TOF MS alone.


2019 ◽  
Vol 36 (No. 6) ◽  
pp. 452-458 ◽  
Author(s):  
Štěpán Koudelka ◽  
Tereza Gelbíčová ◽  
Markéta Procházková ◽  
Renáta Karpíšková

The identification of Listeria species, lineages and serotypes remains a crucial issue not only in epidemic surveys, but also in monitoring of the diversity of bacteria in the food chain. The aim of this study was identification of L. monocytogenes strains at lineage and serotype level using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The performance of MALDI-TOF MS was tested to identify L. monocytogenes into two lineages (I and II) and four serotypes (1/2a, 1/2b, 1/2c and 4b) the most commonly found in humans and food. Total of 227 L. monocytogenes strains from different sources were subjected to the study. Some of strains (112) were used for main spectrum profile (MSP) library creation. Other strains of interest (115) were then correctly identified on the lineage level comparing with the library by MALDI-TOF MS analysis using Biotyper (90%) and ClinPro Tools (100%) software. The serotype identification with 55.7% (Biotyper) and 67.8% (ClinPro Tools) accuracy is rather a proof that under given conditions the method has not big potential to be used for serotyping. However, MALDI-TOF MS has a potential to identify lineages of L. monocytogenes of food and human origin.


2017 ◽  
Vol 55 (4) ◽  
pp. 1162-1176 ◽  
Author(s):  
Andrew M. Borman ◽  
Mark Fraser ◽  
Adrien Szekely ◽  
Daniel E. Larcombe ◽  
Elizabeth M. Johnson

ABSTRACT Exophiala is a ubiquitous pleomorphic genus comprising at least 40 species, many of which have been associated with superficial, visceral, or systemic infections in humans, other mammals, or cold-blooded animals. In this study, we investigated the potential of matrix-assisted laser desorption–ionization time of flight mass spectrometry (MALDI-TOF MS) for the identification of Exophiala species. A total of 89 isolates (including 50 human and 4 animal clinical isolates) stored in the National Collection of Pathogenic Fungi were identified by PCR amplification and sequencing of internal transcribed spacer region 1. Eighty-three of the isolates corresponded to 16 known species within Exophiala/Rhinocladiella . The remaining six isolates are shown by phylogenetic analyses based on four loci to represent two novel Exophiala species. Four isolates from domestic bathrooms which form a sister species with Exophiala lecanii-corni are described here as Exophiala lavatrina sp. nov. The remaining two isolates, both from subcutaneous infections, are distantly related to Exophiala oligosperma and are described here as Exophiala campbellii sp. nov. The triazoles and terbinafine exhibited low MICs against all Exophiala isolates in vitro . MALDI-TOF MS successfully distinguished all 18 species and identified all isolates after appropriate reference spectra were created and added to commercial databases. Intraspecific mean log scores ranged from 1.786 to 2.584 and were consistently significantly higher than interspecific scores (1.193 to 1.624), with the exception of E. lecanii-corni and E. lavatrina , for which there was considerable log score overlap. In summary, MALDI-TOF MS allows the rapid and accurate identification of a wide range of clinically relevant Exophiala species.


2015 ◽  
Vol 54 (2) ◽  
pp. 376-384 ◽  
Author(s):  
S. P. Buckwalter ◽  
S. L. Olson ◽  
B. J. Connelly ◽  
B. C. Lucas ◽  
A. A. Rodning ◽  
...  

The value of matrix-assisted laser desorption ionization−time of flight mass spectrometry (MALDI-TOF MS) for the identification of bacteria and yeasts is well documented in the literature. Its utility for the identification of mycobacteria andNocardiaspp. has also been reported in a limited scope. In this work, we report the specificity of MALDI-TOF MS for the identification of 162Mycobacteriumspecies and subspecies, 53Nocardiaspecies, and 13 genera (totaling 43 species) of other aerobic actinomycetes using both the MALDI-TOF MS manufacturer's supplied database(s) and a custom database generated in our laboratory. The performance of a simplified processing and extraction procedure was also evaluated, and, similar to the results in an earlier literature report, our viability studies confirmed the ability of this process to inactivateMycobacterium tuberculosisprior to analysis. Following library construction and the specificity study, the performance of MALDI-TOF MS was directly compared with that of 16S rRNA gene sequencing for the evaluation of 297 mycobacteria isolates, 148Nocardiaspecies isolates, and 61 other aerobic actinomycetes isolates under routine clinical laboratory working conditions over a 6-month period. MALDI-TOF MS is a valuable tool for the identification of these groups of organisms. Limitations in the databases and in the ability of MALDI-TOF MS to rapidly identify slowly growing mycobacteria are discussed.


2004 ◽  
Vol 48 (2) ◽  
pp. 466-472 ◽  
Author(s):  
Florian Marks ◽  
Christian G. Meyer ◽  
Jürgen Sievertsen ◽  
Christian Timmann ◽  
Jennifer Evans ◽  
...  

ABSTRACT Increasing resistance, recrudescences, and treatment failure have led to the replacement of chloroquine with the combination of pyrimethamine (PYR) and sulfadoxine (SDX) as the first-line antimalarial drugs for treatment of uncomplicated Plasmodium falciparum malaria in several areas where this disease is endemic. The development of resistance to PYR-SDX is favored by incomplete treatment courses or by subtherapeutic levels in plasma. PYR-SDX resistance has been associated with several single-nucleotide polymorphisms (SNPs) in the P. falciparum dihydrofolate reductase (pfdhfr) and the P. falciparum dihydropteroate synthetase (pfdhps) genes. We have established assays based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) that conveniently allow the identification of SNPs associated with PYR resistance. Variants occurring at codon positions 16, 51, 59, and 108 of the pfdhfr gene were analyzed by MALDI-TOF MS in synthetic oligonucleotides to determine the detection threshold. In addition, 63 blood samples from subjects with P. falciparum parasitemia of various degrees were analyzed. The results were compared to those obtained by DNA sequencing of the respective gene fragment. The results of MALDI-TOF MS and DNA sequencing were consistent in 40 samples. In 23 samples two or three pfdhfr variants were detected by MALDI-TOF assays, whereas DNA-sequencing revealed one variant only. Simultaneous detection of two different mutations by biplex assays was, in principle, feasible. As demonstrated by the example of PYR resistance, MALDI-TOF MS allows for rapid and automated high-throughput assessment of drug sensitivity in P. falciparum malaria.


Sign in / Sign up

Export Citation Format

Share Document